MA1-216

antibody from Invitrogen Antibodies

Targeting: THRB ERBA-BETA, ERBA2, GRTH, NR1A2, PRTH, THR1, THRB1, THRB2

Provider product page for MA1-216

Western blot

Immunoprecipitation

Immunohistochemistry

Gel shift

Other assay

Antibody data

Antibody Data
Antigen structure
References [33]
Comments [0]
Validations
Western blot [1]
Immunohistochemistry [2]
Other assay [10]

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Product number: MA1-216 - Provider product page
Provider: Invitrogen Antibodies
Product name: THRB Monoclonal Antibody (J52)
Antibody type: Monoclonal
Antigen: Purifed from natural sources
Reactivity: Human, Mouse, Rat
Host: Mouse
Isotype: IgG
Antibody clone number: J52
Vial size: 100 µL
Concentration: 1 mg/mL
Storage: -20° C, Avoid Freeze/Thaw Cycles

THRB protein structure - MA1-216 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry was performed on normal biopsies of deparaffinized Human thyroid tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature and probed with a Thyroid Hormone Receptor beta-1 monoclonal antibody (Product # MA1-216) at a dilution of 1:20 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of Thyroid Hormone Receptor beta-1 showing staining in the nucleus and weak staining in the cytoplasm of paraffin-embedded human colon tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Thyroid Hormone Receptor beta-1 Mouse Monoclonal Antibody (Product # MA1-216) diluted in 3% BSA-PBS at a dilution of 1:20 for 1 hour at 37ºC in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 4 Characterization of TRbeta binding near induced genes. A. Patterns of TRbeta binding depicted at representations of individual target gene loci (LDLR, BCL3, NCOR2,ADSSL1 and SOX7). Blue bars represent genomic binding regions, and the vertical red lines represent peaks, as classified by QuEST. The horizontal black bars are regions analyzed by ChIP-PCR (locations of primer amplification). Observed binding patterns included 5', 3' and intronic binding events, as shown in genomic data tracks (UCSC Genome Browser). Putative regulatory elements, identified through sequence analysis of the genomic regions indicated, are depicted below bound regions in which they occur. B. QPCR of ChIP analysis confirming DNA binding in regulatory regions of genes. C. Realtime PCR analysis depicting enhancement of transcription of individual loci in Fig. 4A by T3 in the presence of TRbeta. (*P

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 6 Links between TR Binding and Adm Transcription. A. Graph showing results of realtime PCR analysis of adm transcription in the B7B cells after six hours of T3 treatment +/-10 ug/ml CHX cotreatment of B7B cells. B. Patterns of TRbeta binding peaks at the adm locus (UCSC Genome Browser), in similar format to Fig. 4 . TRbeta binding events clustered into four regions (R1, R2, R3, R4), upstream and downstream of this transcript, as well as a substantial amount of binding immediately proximal to the transcriptional start site. C. Binding of TRbeta was confirmed by realtime ChIP PCR analysis in B7B cells at the regions indicated (ChIP primers are depicted by horizontal bars in B). D. The proximal promoter region of adm (corresponding to R2) conferred T3-dependent increases in luciferase activity upon a standard reporter after transfection into B7B. E. Results of gel shift confirming direct TRbeta binding to two putative response elements, designated TRE-1 and TRE-2 that were found in R2 at positions marked in Fig. 6B . Individual lanes show shifts obtained with elements and RXRalpha-TRbeta +/- competitor DNA or mutated versions of both elements. F. Luciferase reporter assays confirming that TRE-1 and TRE-2 confer T3 responsiveness on a reporter gene. (*P

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 7 Intergenic binding events. A. Three intergenic binding peaks were selected and analyzed for the presence of recognizable TR binding motifs (sequences of motifs listed). B. Results of qPCR ChIP analysis confirming binding of TRbeta to the intergenic regions depicted in Fig. 7A (top panels) and induction of H3 acetylation near sites (bottom panels). C. Results of luciferase reporter assays, with indicated constructs containing intergenic elements described in Fig. 7A , confirming that each element confers T3 induction. (*P

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 10 Effects of TH replacement on TH-related proteins in THRSP OE mice. The TH-related proteins were evaluated following 7 days of LT3 and LT4 (10 mg kg -1 ) treatment. On the last day of the treatment, the brains were harvested, and the striatum was subjected to western blot analysis. a Representative blots for specific protein targets. b MCT8, c TRalpha, and d TRbeta protein levels ( n = 6 mice/group; b two-way ANOVA, F (1,30) = 4.96, P = 0.034; c two-way ANOVA, F (1,30) = 14.4, P < 0.001; d two-way ANOVA, F (1,30) = 0.123, P = 0.728). TH replacement improved the striatal MCT8 and TRalpha levels in THRSP OE mice. Values are presented as the mean +- S.E.M. * P < 0.05 (relative to WT) and # P < 0.05. ## P < 0.01 (relative to VEH treatment), by two-way ANOVA with Bonferroni's multiple comparison tests.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 2 Reduction of TRbeta1 in Tg197 LV samples and female-specific reduction of Akt activation. Densitometric assessment in arbitrary units and representative Western blots of thyroid hormone receptor alpha1 nuclear expression (TRalpha1, A ), thyroid hormone receptor beta1 nuclear expression (TRbeta1, B ) and phosphorylated and total p38 MAPK ( C ) in left ventricle of WTC57BL/6JxCBA (WT, n = 4 per gender) and Tg197 ( n = 6 per gender) male and female mice. Values are represented in mean +- SEM; * p < 0.05 (H3: Histone 3).